Anticancer potential of isolated fractions of muntingia calabura l.using HT-29 cell line

Abstract

Cancer is a large family of diseases that involve abnormal cell growth with the potential to invade or spread to the other parts. A neoplasm or tumor is a group of cells that have undergone unregulated growth and will often form a mass or lump, but be distributed diffusely. To extract the leaves of Muntingia calibra L. using chloroform by cold maceration method. To isolate the phytoconstituents with column chromatography technique, and evaluate the anticancer activity of isolated fractions on HT-29 cell line using MTT assay method. Perturbing factor must first be investigated via a cell viability assay. The viability assay most commonly used throughout the world is the MTT assay, first described by Tim Mossman in 1983. This colorimetric assay uses reduction of a yellow tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide, or MTT) to measure cellular metabolic activity as a proxy for cell viability. Viable cells contain NAD(P)H-dependent oxidoreductase enzymes which reduce the MTT reagent to formazan, an insoluble crystalline product with a deep purple color. Formazan crystals are then dissolved using a solubilising solution and absorbance is measured at 500- 600 nanometers using a plate-reader. The darker the solution, the greater the number of viable, metabolically active cells. Each condition should be done in triplicate or more. Phytochemical constituents of carbohydrates, proteins, glycosides, cardiacglycosides terpenoids, saponins, flavonoids and phenolic compound are present in the chloroform leaf extract. Three compounds re isolated from chloroform leaf extract of Muntingia calubra L. by using column chromatography. The isolated fractions F3M, F5MF8M have anticancer activity among these fractions F3M have more potent anti-cancer activity on HT-29 cell lines.